Liu Changqi, Chhabra Guneet S, Zhao Jing, Zaffran Valerie D, Gupta Sahil, Roux Kenneth H, Gradziel Thomas M, Sathe Shridhar K
Dept. of Nutrition, Food & Exercise Sciences, Florida State Univ., Tallahassee, FL, 32306-1490, U.S.A.
Dept. of Biological Science, Florida State Univ., Tallahassee, FL, 32306-4370, U.S.A.
J Food Sci. 2017 Oct;82(10):2504-2515. doi: 10.1111/1750-3841.13829. Epub 2017 Sep 4.
A commercially available monoclonal antibody (mAb)-based direct sandwich enzyme-linked immunosorbent assay (ELISA) kit (BioFront Technologies, Tallahassee, Fla., U.S.A.) was compared with an in-house developed mAb 4C10-based ELISA for almond detection. The assays were comparable in sensitivity (limit of detection < 1 ppm full fat almond, limit of quantification < 5 ppm full fat almond), specificity (no cross-reactivity with 156 tested foods at a concentration of 100000 ppm whole sample), and reproducibility (intra- and interassay variability < 15% CV). The target antigens were stable and detectable in whole almond seeds subjected to autoclaving, blanching, frying, microwaving, and dry roasting. The almond recovery ranges for spiked food matrices were 84.3% to 124.6% for 4C10 ELISA and 81.2% to 127.4% for MonoTrace ELISA. The almond recovery ranges for commercial and laboratory prepared foods with declared/known almond amount were 30.9% to 161.2% for 4C10 ELISA and 38.1% to 207.6% for MonoTrace ELISA. Neither assay registered any false-positive or negative results among the tested commercial and laboratory prepared samples.
Ability to detect and quantify trace amounts of almonds is important for improving safety of almond sensitive consumers. Two monoclonal antibody-based ELISAs were compared for almond detection. The information is useful to food industry, regulatory agencies, scientific community, and almond consumers.
将一种市售的基于单克隆抗体（mAb）的直接夹心酶联免疫吸附测定（ELISA）试剂盒（美国佛罗里达州塔拉哈西市的BioFront Technologies公司）与一种内部开发的基于mAb 4C10的ELISA方法用于杏仁检测进行了比较。这两种检测方法在灵敏度（检测限＜1 ppm全脂杏仁，定量限＜5 ppm全脂杏仁）、特异性（在100000 ppm全样品浓度下与156种受试食品无交叉反应）和重现性（批内和批间变异系数＜15%）方面具有可比性。目标抗原在经过高压灭菌、去皮、油炸、微波处理和干烤的完整杏仁种子中稳定且可检测。对于加标食品基质，4C10 ELISA的杏仁回收率范围为84.3%至124.6%，MonoTrace ELISA为81.2%至127.4%。对于已申报/已知杏仁含量的市售和实验室制备食品，4C10 ELISA的杏仁回收率范围为30.9%至161.2%，MonoTrace ELISA为38.1%至207.6%。在受试的市售和实验室制备样品中，两种检测方法均未出现假阳性或假阴性结果。
检测和定量痕量杏仁的能力对于提高杏仁敏感消费者的安全性很重要。比较了两种基于单克隆抗体的ELISA方法用于杏仁检测。该信息对食品行业、监管机构、科学界和杏仁消费者有用。
